Abstracts / Atherosclerosis 233 (2014) 326–330 327
EXPRESSION OF APOESENDAI BY MACROPHAGES CAUSES LIPOPROTEINGLOMERULOPATHY WHILE SUPPRESSING ATHEROSCLEROSIS
Hagai Tavori a, Daping Fan d, Shilin Yang a, Agnes B. Fogo c, MacRae F.Linton a,b, Sergio Fazio a,c
a Section of Cardiovascular Disease Prevention, Department of Medicine,Vanderbilt University Medical Center, Nashville, TN; bDepartment ofPharmacology, Vanderbilt University Medical Center, Nashville, TN;cDepartment of Pathology, Microbiology and Immunology, VanderbiltUniversity Medical Center, Nashville, TN; dDepartment of Cell Biology andAnatomy, University of South Carolina, Columbia, SC, USA
Renal disease is usually associated with lipoprotein abnormalities. Incontrast, lipoprotein glomerulopathy (LPG) is a unique disease character-ized by intraglomerular lipoprotein thrombi, often in the absence of dys-lipidemia. Despite the glomerular lipid accumulation, macrophages areapparently not involved in LPG. ApoESendai is an apoE variant known tocause LPG in heterozygous carriers. It carries the same charge as the apoE2common variant, but is due to a proline-for-arginine substitution at po-sition 145 in the apoE3 sequence.We have characterized plasma lipids and renal and arterial responses inchimericapoE�/�orapoE�/�/LDLR�/�doubleknockout (DKO)miceexpressingapoESendai exclusively in macrophages. Bone marrow from apoE�/� mice wastransducedwith lentiviral constructsexpressingeitherhumanapoE2 (control)or apoESendai, and then transplanted into apoE�/� or DKO recipient mice.Compared to BMT controls, both apoESendai and apoE2 macrophages signifi-cantly decreased plasma cholesterol levels in apoE�/� recipients (28% and13%, respectively). No changes in serum cholesterol levelswere found inDKOrecipients compared to controls. Macrophage expression of apoESendai andapoE2 was associated with plasma human apoE levels of 6.9 � 1.9 and 5.4 �1.7mg/ml, respectively, in apoE�/� recipients, and18.9� 2.8 and18.4� 3.8mg/ml, respectively, inDKO recipients.Macrophage apoE2expressionbyapoE�/�
BMT recipients reduced aortic lesion size by 23% compare to BMTcontrols (p< 0.01). Surprisingly, macrophage apoESendai expression further reducedaortic lesionsizeby33.7%comparedtoapoE2expression inapoE�/�mice (p<0.01). A similar 26.1% reductionwas seen inDKO recipients transplantedwithapoESendai compared to apoE2 expressing macrophages.Kidneys had glomerular lesions and vacuoles in the proximal tubules,which correlates with the severity of dyslipidemia in all mice, in contrastonly DKO mice expressing apoESendai showed focal mesangiolysis withmoderate to severe increase in mesangial matrix, clear features of LPG.Our results show for the first time that macrophage expression ofapoESendai protects against atherosclerosis while causing glomerular disease.
ASSESSING THE TREATMENT EFFECT IN METABOLIC SYNDROMEWITHOUT PERCEPTIBLE DIABETES (ATTEMPT): A PROSPECTIVE-RANDOMIZED STUDY IN MIDDLE AGED MEN AND WOMEN
V.G. Athyros, E. Ganotakis, G.D. Kolovou, V. Nicolaou, A. Achimastos, E.Bilianou, T. Alexandrides, A. Karagiannis, K. Paletas, E.N. Liberopoulos, K.Tziomalos, D. Petridis, A. Kakafika, M.S. Elisaf, D.P. Mikhailidis
Assessing The Treatment Effect in Metabolic Syndrome Without PerceptibleDiabetes (ATTEMPT) Collaborative Hellenic Atherosclerosis Society
Aim: To assess the reduction in estimated cardiovascular disease (e-CVD)risk after multifactorial treatment for 6 months and follow this changeduring the next 3 years.Patients and methods: This prospective, randomized, target-driven studyincluded 1,123 subjects (512 men, aged 45–65 years) with metabolicsyndrome (MetS) without diabetes or CVD referred to specialist outpatientclinics. Patients were randomized to two treatment groups: group A withlow density lipoprotein cholesterol (LDL-C) target of <100 mg/dl andgroup B with a target of <130 mg/dl. Atorvastatin was used in both groupson top of optimal multifactorial treatment (quinapril, amlodipine, hydro-chlorothiazide for hypertension, metformin for impaired fasting glucose,and orlistat for obesity). The e-CVD risk was calculated using the Fra-mingham, PROCAM and Reynold's equations.
Results: Reductions in e-CVD risk at 6monthswere>50% in all patients, butwere superior in group A and in women. Reductions were even greaterduring the next 3-years and were mainly attributed to changes in the lipidprofile. Actual CVD events were 1 in group A and 13 in group B (p¼ 0.0012).Conclusions: Attaining the treatment target of LDL-C <100 mg/dl withinmultifactorial treatment of MetS by expert clinics is achievable andbeneficial even in patients without diabetes or known CVD. This induces aconsiderable e-CVD risk reduction in MetS patients. Actual CVD eventswere negligible, suggesting that e-CVD risk overestimates actual CVD riskin MetS, at least in patients achieving LDL-C <100 mg/dl.
CRP EXPRESSION AND MODULATION IN DIFFERENT SUBTYPES OFMACROPHAGES: IMPLICATIONS IN THE PATHOGENESIS OFATHEROGENESIS
Shur Anna, Tendler Yevgeny, Kaplan Marielle
Laboratory of Clinical Biochemistry, RambamHealth Care Campus, Haifa, Israel
Atherosclerotic plaque residents –macrophages, that are key players in theinflammatory processes in the artery wall, comprise a heterogeneouspopulation, including several subtypes: pro-inflammatory (M1) and anti-inflammatory (M2). According to recent studies, C Reactive protein (CRP) ispresent in the arterial intima of atherosclerotic lesions. CRP locally pro-duced by macrophages could directly participate in atherogenesis andother cardiovascular complications.The primary aim of this study was to develop a system for induction ofmacrophages toM1 andM2 phenotypes. In addition, we analyzed whetherthere were differences in CRP expression in M1 versus M2 macrophages.PMA-induced Thp-1 macrophages were used in this study. M1 phenotypeinduction was performed by cells incubation with INF-g and LPS, whereasM2 phenotype induction was performed by cells incubation with IL-4. M1and M2 phenotype were assessed by measuring IL-6 and IL-10 levels inculture media, respectively. In addition, M2 phenotype was further char-acterized by measuring the level of expression of mannose receptor usingFACS technique. CRP mRNA levels were measured both in M1/M2 cells byreal time PCR. Moreover, intracellular levels of CRP protein were deter-mined by immunohistochemistry staining.Following concentrations-dependent studies, M1 phenotype differentia-tion was validated by macrophages incubation with 60 ng/ml IFN-g in thepresence of 100 ng/ml LPS. Moreover, M2 phenotype differentiation wasvalidated following macrophages incubation with 60 ng/ml IL-4. M1 pro-inflammatory phenotype showed significant increase in CRP mRNAexpression by 68% and intracellular level of CRP protein by 108%. M2phenotype exhibited no change in CRP expression (mRNA and protein) incomparison to control untreated cells.Our data supports that elevated expression of CRP is characteristic of M1pro-inflammatory macrophages, whereas induction of macrophages toM2phenotypes does not induce CRP cellular expression. Therefore, switchingthe induction of macrophages from M1 pathway to M2 pathway couldreduce CRP deleterious effects as well as the inflammatory cascade in theatherosclerotic plaque.
A PRACTICAL METHOD TO ASSESS APOLIPOPROTEIN PROFILES INNATIVE LDL AND HDL SUBCLASSES: A PILOT STUDY
Teresita Menini, Alejandro Gugliucci
Glycation, Oxidation and Disease Laboratory, College of OsteopathicMedicine, Touro University-California, Vallejo, USA
Background: The HDL-C hypothesis has been challenged by recent men-delian randomization studies. Much more information can be obtained byanalyzing the individual apolipoproteins across the spectrum of particles.There are only a few studies using elaborate MALDI-MS proteomics on HDLpurified by ultracentrifugation.Aims of this study: We have developed a practical method to detect themajor apolipoproteins in LDL and HDL subclasses in human serum.
